Ab184951

Reagent grade ICA (purity ≥ 98% by HPLC analysis) was obtained from Nanjing Zelang Medical Technology Corporation Ltd. (Nanjing, China) and dissolved in dimethyl sulfoxide (DMSO); the final concentration of DMSO in the media was less than 0.1% (v/v). NaN₃ (A0639) was purchased from Amresco (Solon, OH, USA). RIPA buffer (high) (R0010) and protein phosphatase inhibitor (P1260) were purchased from Solarbio Life Science (Beijing, China). A glucose oxidase assay kit (E10160) and antibodies against GSK-3β (9315), p-Ser9-GSK-3β (9323) and p-PI3K (p85) (4228) were obtained from Cell Signaling Technology (Beverly, MA, USA). Goat anti-mouse IgG-HRP (SA00001-1), goat anti-rabbit IgG-HRP (SA00001-2), and antibodies against GAPDH (60004-1-Ig), and PI3K p110α (21890-1-AP) were obtained from Proteintech Group (Wuhan, China). Antibodies against PHF1 (ab184951) and PI3K (ab86714) were obtained from Abcam (Cambridge, MA, USA). PageRuler prestained protein ladder (26616) and antibodies against p-T217 (44-744) and TAU-5 (MA5-12808) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). RPMI 1640 HyClone™ cell culture medium (SH30809.01) was purchased from GE Healthcare (Chicago, IL, USA).

Antibodies used in the study include: rabbit monoclonal anti-β III tubulin for Western blot analysis (Abcam; ab52901; 1/8,000); rabbit monoclonal anti-PHF1 (Abcam; ab184951; 1/5,000); rabbit monoclonal anti-tau (phospho T231) (Abcam; ab151559; 1/5,000); chickenanti-tau (Abcam; ab75714; 1/30000); rabbit monoclonal anti-mTOR (phospho S2448) (Abcam; ab109268; 1/5,000); rabbit anti-mTOR (Cell Signalling; 2983; 1/1,000); rabbit anti-Phospho-S6K (Thr421/Ser424) (Cell Signalling; 9204; 1/1,000); rabbit anti-S6K (Cell Signalling; 9202; 1/1,000); rabbit anti-Cav-1 for western blot analysis (Santa Cruz Biotechnology; sc-894; 1/500); rabbit anti-Cav-1 for immunohistochemical staining (Santa Cruz Biotechnology; sc-894; 1/100); rabbit anti-Cav-1 for immunofluorescence (Santa Cruz Biotechnology; sc-894; 1/50); rabbit monoclonal anti-Synaptophysin (Syn) (Abcam; ab32127; 1/50); rabbit monoclonal anti-β III tubulin for immunofluorescence (Abcam; ab52901; 1/100).
The adenoviral Cav-1 (Ad-Cav-1) and the empty adenovirus vector (Ad-null) were generously gifted from Dr. Duan-Fang Liao (Hunan University of Traditional Chinese Medicine, China).

The following antibodies were used: rabbit monoclonal anti-GAPDH (Beyotime; AF1186; 1/1000); rabbit monoclonal anti-PHF1 (Abcam; ab184951; 1/1000); rabbit monoclonal anti-tau (phospho-T231; Abcam; ab151559; 1/1000); anti-tau (Affinity; AF6141; 1/1000); rabbit polyclonal anti-ALKBH5 (Proteintech; 16837-1-AP; 1/2000); mouse monoclonal anti-FTO (Abcam; ab92821; 1/1000); rabbit polyclonal anti-METTL3 (Proteintech; 15073-1-AP; 1/1000); rabbit polyclonal anti-METTL14 (ABclonal; A8530; 1/1000); rabbit polyclonal anti-Dgkh (Proteintech; 13873-1-AP; 1/1000); and anti-m⁶A (Synaptic Systems; 202,003).